Prostate-Specific Membrane Antigen Binding Proteins and Related Compositions and Methods

Abstract

The present invention relates to mono-specific and multi-specific polypeptide therapeutics that specifically target cells expressing prostate-specific membrane antigen (PSMA) and are useful for the treatment of prostate cancer (e.g., castrate-resistant prostate cancer), tumor-related angiogenesis or benign prostatic hyperplasia (BPH). In one embodiment, the multi-specific polypeptide therapeutics bind both PSMA-expressing cells and the T-cell receptor complex on T cells to induce target-dependent T-cell cytotoxicity, activation and proliferation.

1 .- 153 . (canceled) 
     
     
         154 . A prostate-specific membrane antigen (PSMA)-binding polypeptide comprising a humanized PSMA-binding domain wherein the PSMA binding domain comprises:
 (i) an immunoglobulin light chain variable region comprising LCDR1, LCDR2, and LCDR3, and   (ii) an immunoglobulin heavy chain variable region comprising HCDR1, HCDR2, and HCDR3,   wherein the LCDR1, LCDR2 and LCDR3 has the amino acid sequences set forth in SEQ ID NOs: 15, 16 and 17, respectively, and the HCDR1, HCDR2, and HCDR3 has the amino acid sequences set forth in SEQ ID NOs: 9, 10 and 11, respectively.   
     
     
         155 . The PSMA-binding polypeptide of  claim 154 , further comprising a first hinge region. 
     
     
         156 . The PSMA-binding polypeptide of  claim 155 , wherein the first hinge region is derived from (i) a stalk region of a type II C lectin or (ii) an immunoglobulin hinge region. 
     
     
         157 . The PSMA-binding polypeptide of  claim 155 , further comprising an immunoglobulin constant region. 
     
     
         158 . The PSMA-binding polypeptide of  claim 157 , wherein the immunoglobulin constant region comprises immunoglobulin CH2 and CH3 domains of IgG1, IgG2, IgG3, IgG4, IgA1, IgA2 or IgD. 
     
     
         159 . The PSMA-binding polypeptide of  claim 159 , wherein the PSMA-binding polypeptide comprises at least one effector function selected from the group consisting of antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). 
     
     
         160 . The PSMA-binding polypeptide of  claim 157 , wherein said PSMA-binding polypeptide comprises from amino-terminus to carboxyl-terminus
 (a) the PSMA binding domain,   (b) the first hinge region, and   (c) the immunoglobulin constant region.   
     
     
         161 . The PSMA-binding polypeptide of  claim 154 , wherein said PSMA-binding polypeptide comprises an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:70, or SEQ ID NO:72. 
     
     
         162 . The PSMA-binding polypeptide of  claim 154 , wherein said PSMA-binding polypeptide further comprises a second binding domain. 
     
     
         163 . The PSMA-binding polypeptide of  claim 162 , wherein the PSMA-binding polypeptide comprises, in order from amino-terminus to carboxyl-terminus or in order from carboxyl-terminus to amino-terminus,
 (a) the PSMA binding domain,   (b) a first hinge region,   (c) an immunoglobulin constant region,   (d) a second hinge region, and   (e) the second binding domain.   
     
     
         164 . The PSMA-binding polypeptide of  claim 163 , wherein the second hinge region is derived from (i) a stalk region of a type II C lectin or (ii) an immunoglobulin hinge region. 
     
     
         165 . The PSMA-binding polypeptide of  claim 162 , wherein the second binding domain specifically binds a T cell, CD3, CD3ε or a T cell receptor (TCR) complex or a component thereof. 
     
     
         166 . The PSMA-binding polypeptide of  claim 162 , wherein the second binding domain competes for binding to CD3ε with a monoclonal antibody selected from the group consisting of CRIS-7 and HuM291. 
     
     
         167 . The PSMA-binding polypeptide of  claim 162 , wherein the second binding domain comprises an immunoglobulin light chain variable region and an immunoglobulin heavy chain variable region derived from a monoclonal antibody selected from the group consisting of CRIS-7 and HuM291. 
     
     
         168 . The PSMA-binding polypeptide of  claim 167 , wherein the light and heavy chain variable regions of the second binding domain are humanized variable regions of the light and heavy chain CDRs of the monoclonal antibody. 
     
     
         169 . The PSMA-binding polypeptide of  claim 167 , wherein the light and heavy chain variable regions of the second binding domain are selected from the group consisting of:
 (a) a light chain variable region comprising an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in residues 139-245 of SEQ ID NO:47 and a heavy chain variable region comprising an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in residues 1-121 of SEQ ID NO:47; and   (b) a light chain variable region comprising an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in residues 634-740 of SEQ ID NO:78 and a heavy chain variable region comprising an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in residues 496-616 of SEQ ID NO:78.   
     
     
         170 . The PSMA-binding polypeptide of  claim 162 , wherein the second binding domain is a single chain Fv (scFv). 
     
     
         171 . The PSMA-binding polypeptide of  claim 154 , wherein said PSMA-binding polypeptide comprises an amino acid sequence that is at least 95% or 100% identical to the amino acid sequence set forth in SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:74, SEQ ID NO:76, SEQ ID NO:78, SEQ ID NO:80, SEQ ID NO:82, SEQ ID NO:84, SEQ ID NO:158, SEQ ID NO:160, SEQ ID NO:162, or SEQ ID NO:164. 
     
     
         172 . The PSMA-binding polypeptide of  claim 154 , wherein the immunoglobulin light chain variable region comprises an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO:5 or SEQ ID NO:23 and the heavy chain variable region comprises an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO:2, SEQ ID NO:25, or SEQ ID NO:27. 
     
     
         173 . The PSMA-binding polypeptide of  claim 172 , wherein the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO:23 and the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO:25 or SEQ ID NO:27. 
     
     
         174 . The PSMA-binding polypeptide of  claim 154 , wherein the PSMA-binding domain competes for binding to human PSMA with a single chain Fv (scFv) having the amino acid sequence set forth in SEQ ID NO:21. 
     
     
         175 . The PSMA-binding polypeptide of  claim 154 , wherein the PSMA-binding domain is a single chain Fv (scFv). 
     
     
         176 . The PSMA-binding polypeptide of  claim 175 , wherein the light chain variable region and heavy chain variable region of the scFv are joined by an amino acid sequence comprising (Gly 4 Ser) n , wherein n=1-5 (SEQ ID NO: 165). 
     
     
         177 . The PSMA-binding polypeptide of  claim 175 , wherein the scFv comprises an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:34, or SEQ ID NO:35. 
     
     
         178 . The PSMA-binding polypeptide of  claim 154 , further comprising an immunoglobulin heterodimerization domain. 
     
     
         179 . The PSMA-binding polypeptide of  claim 178 , wherein the immunoglobulin heterodimerization domain comprises an immunoglobulin CH1 domain or an immunoglobulin CL domain. 
     
     
         180 . The PSMA-binding polypeptide of  claim 178 , wherein said PSMA-binding polypeptide comprises an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, or SEQ ID NO:61. 
     
     
         181 . A dimeric PSMA-binding protein comprising first and second polypeptide chains, wherein each of said polypeptide chains is the PSMA-binding polypeptide of  claim 154 . 
     
     
         182 . A composition comprising the PSMA-binding protein of  claim 181  and a pharmaceutically acceptable carrier, diluent, or excipient. 
     
     
         183 . An isolated nucleic acid encoding the PSMA-binding polypeptide of  claim 154 . 
     
     
         184 . A recombinant host cell comprising the nucleic acid of  claim 183 . 
     
     
         185 . A method for producing a dimeric PSMA-binding protein, the method comprising: culturing a recombinant host cell comprising an expression vector, wherein the expression vector comprises a nucleic acid segment encoding the PSMA-binding polypeptide of  claim 154 , wherein the nucleic acid segment is operably linked to regulatory sequences suitable for expression of the PSMA-binding polypeptide in the host cell, and wherein said culturing is under conditions whereby the PSMA-binding polypeptide is expressed and produced as a dimeric PSMA-binding protein. 
     
     
         186 . A method for inducing at least one of antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) against a cell expressing prostate-specific membrane antigen (PSMA), the method comprising: contacting said PSMA-expressing cell with a dimeric PSMA-binding protein comprising first and second polypeptide chains, wherein each of said polypeptide chains is the PSMA-binding polypeptide of  claim 159 , wherein said contacting is under conditions whereby at least one of ADCC and CDC against the PSMA-expressing cell is induced. 
     
     
         187 . A method for inducing redirected T-cell cytotoxicity (RTCC) against a cell expressing prostate-specific membrane antigen (PSMA), the method comprising contacting said PSMA-expressing cell with the PSMA-binding protein of  claim 165 , wherein said contacting is under conditions whereby RTCC against the PSMA-expressing cell is induced. 
     
     
         188 . A method for treating a disorder in a subject, wherein said disorder is characterized by overexpression of PSMA, the method comprising administering to the subject a therapeutically effective amount of the dimeric PSMA-binding protein of  claim 181 . 
     
     
         189 . The method of  claim 188 , wherein the disorder is a cancer, a prostate disorder, or a neovascular disorder. 
     
     
         190 . The method of  claim 188 , wherein the disorder is prostate cancer, colorectal cancer, gastric cancer, castrate-resistant prostate cancer, benign prostatic hyperplasia, solid tumor growth, clear cell renal carcinoma, colorectal cancer, bladder cancer, or lung cancer. 
     
     
         191 . A chimeric or humanized prostate-specific membrane antigen (PSMA)-binding polypeptide comprising
 a) a PSMA binding domain that specifically binds human PSMA,   (b) a hinge region, and   (c) an immunoglobulin constant region.   
     
     
         192 . A heterodimeric PSMA-binding protein comprising
 (1) a first polypeptide chain comprising, in order from amino-terminus to carboxyl-terminus,
 (a) a PSMA binding domain that specifically binds human PSMA, 
 (b) a first hinge region, 
 (c) a first immunoglobulin constant region, and 
 (d) a first immunoglobulin heterodimerization domain; and 
   (2) a second polypeptide chain comprising, in order from amino-terminus to carboxyl-terminus,
 (a′) a second hinge region, 
 (b′) a second immunoglobulin constant region, and 
 (c′) a second immunoglobulin heterodimerization domain that is different from the first immunoglobulin heterodimerization domain of the first single chain polypeptide, 
 wherein the first and second immunoglobulin heterodimerization domains associate with each other to form a heterodimer. 
   
     
     
         193 . The heterodimeric PSMA-binding protein of  claim 192 , wherein the first immunoglobulin heterodimerization domain comprises an immunoglobulin CH1 domain and the second immunoglobulin heterodimerization domain comprises an immunoglobulin CL domain, or wherein the first immunoglobulin heterodimerization domain comprises an immunoglobulin CL domain and the second immunoglobulin heterodimerization domain comprises an immunoglobulin CH1 domain. 
     
     
         194 . The heterodimeric PSMA-binding protein of  claim 192 , wherein at least one of the first and second immunoglobulin constant regions comprises immunoglobulin CH2 and CH3 domains of IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, IgD or any combination thereof; an immunoglobulin CH3 domain of IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, IgD, IgE, IgM or any combination thereof; or immunoglobulin CH3 and CH4 domains of IgE, IgM or a combination thereof. 
     
     
         195 . The heterodimeric PSMA-binding protein of  claim 192 , wherein the heterodimeric PSMA-binding polypeptide comprises at least one effector function selected from the group consisting of antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). 
     
     
         196 . The heterodimeric PSMA-binding protein of  claim 192 , wherein said second polypeptide chain further comprises a second binding domain. 
     
     
         197 . The heterodimeric PSMA-binding protein of  claim 196 , wherein the second binding domain is amino-terminal to the second hinge region. 
     
     
         198 . The heterodimeric PSMA-binding protein of  claim 192 , wherein the PSMA binding domain comprises (i) an immunoglobulin light chain variable region comprising LCDR1, LCDR2, and LCDR3, and (ii) an immunoglobulin heavy chain variable region comprising HCDR1, HCDR2, and HCDR3, wherein the LCDR1, LCDR2 and LCDR3 has the amino acid sequences set forth in SEQ ID NO: 15, 16 and 17, respectively, and the HCDR1, HCDR2, and HCDR3 has the amino acid sequences set forth in SEQ ID NO:9, 10 and 11, respectively. 
     
     
         199 . The heterodimeric PSMA-binding protein of  claim 192 , wherein
 (a) the first polypeptide chain comprises an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO: 46 and the second polypeptide chain comprises an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO: 47;   (b) the first polypeptide chain comprises an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO: 58 and the second polypeptide chain comprises an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO: 57;   (c) the first polypeptide chain comprises an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO: 59 and the second polypeptide chain comprises an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO: 57;   (d) the first polypeptide chain comprises an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO: 60 and the second polypeptide chain comprises an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO: 47; or   (e) the first polypeptide chain comprises an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO: 61 and the second polypeptide chain comprises an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO: 47.   
     
     
         200 . An expression vector comprising first and second expression units, wherein the first and second expression units respectively comprise first and second nucleic acid segments encoding the first and second polypeptide chains of the heterodimeric PSMA-binding protein of  claim 192 , and wherein the first and second nucleic acid segments are operably linked to regulatory sequences suitable for expression of the nucleic acid segments in a host cell. 
     
     
         201 . A method for producing a heterodimeric PSMA-binding protein, the method comprising: culturing a recombinant host cell comprising first and second expression units, wherein the first and second expression units respectively comprise first and second nucleic acid segments encoding the first and second polypeptide chains of the heterodimeric PSMA-binding protein of  claim 192 , wherein the first and second nucleic acid segments are operably linked to regulatory sequences suitable for expression of the heterodimeric PSMA-binding protein in a host cell, and
 wherein said culturing is under conditions whereby the first and second polypeptide chains of a heterodimeric PSMA-binding protein are expressed and the encoded polypeptide chains are produced as the heterodimeric PSMA-binding protein   
     
     
         202 . A composition comprising the heterodimeric PSMA-binding protein of  claim 192  and a pharmaceutically acceptable carrier, diluent, or excipient. 
     
     
         203 . A method for inducing at least one of antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) against a cell expressing prostate-specific membrane antigen (PSMA), the method comprising: contacting said PSMA-expressing cell with the heterodimeric PSMA-binding protein of  claim 195 , wherein said contacting is under conditions whereby at least one of ADCC and CDC against the PSMA-expressing cell is induced 
     
     
         204 . A method for treating a disorder in a subject, wherein said disorder is characterized by overexpression of PSMA, the method comprising administering to the subject a therapeutically effective amount of the heterodimeric PSMA-binding protein of  claim 192 .